The present invention relates to an apparatus for real-time monitoring chemical reaction between various biomaterials. More particularly, the present invention is directed to a real-time monitoring apparatus for biochemical reaction, which comprises a temperature control block system comprising a thermoelectric element, capable of supplying heat into a reaction tube, a heat transmission block which transmit the heat to the reaction tube; a light irradiation source part comprising a lamp which irradiates light with uniform intensity on sample contained in the reaction tube, and optical waveguide; and optical system comprising a light receiving part for receiving fluorescence irradiated from the sample by the light emitted from the light irradiation source.
Recently, the research and development for chemical microprocessor have been performed actively, which can carry out pretreatment of sample, reaction, separation, detection and etc., within a single chip, so called lab-on-a-chip. The lab-on-a-chip composed from glass, silicon or plastic material and manufactured through the lithography technology which has been employed for semiconductor chip, mounts micro-sized device for analyzing samples rapidly and sensitively.
The above all procedures required for the analysis of sample, i.e., pretreatment, reaction, separation, detection and etc., can be performed continuously. In addition, by using the chip the time needed for the sample analysis can be reduced to second or minute level and also the amount of sample can be reduced to micro-liter level and the size of apparatus on which the chip is mounted can also be miniaturized. The lab-on-a-chip technology is based on the capillary electrophoresis developed by Harrison in early 1990s, and had been started to be known to the public by the fact that a small size lab. Device for the capillary electrophoresis analysis can be integrated into a single chip.
Meanwhile, recently, so called real-time PCR technology which can check rapidly the progress of every cycle of amplification reaction by detection of fluorescence from reaction tubes, without using the separation step in gel phase. The conventional apparatus employed for this real-time PCR technology may be manufactured by coming the thermal cycler for PCR and the fluorometer for detection of products.
The conventional real-time PCR apparatus is composed of the thermoelectric element, the heat transmitting block which transmit heat into reaction tubes which contains sample, the light irradiation source which irradiate light into sample contained in the reaction tube, and the light receiving part which accept the fluorescence generated from the sample.
By using the conventional real-time PCR apparatus, the progress of PCR can be checked in real-time by measuring the intensity of fluorescence generated from sample upon completion of each cycle by the operation of the thermoelectric element for cooling and heating repeatedly to carry out the reaction of biochemical sample contained in the tube.
In the conventional real-time PCR apparatus, in general, a halogen lamp and a metallic halide lamp have been employed as a light irradiation source. In this conventional apparatus, a selective transmission filter(9) transmit selectively the light with preferred wavelength from the light radiated from the lamp(5) and then, the light thus selected irradiated into the sample contained in the tubes via a reflection mirror(18) and through a condensing lens(17).
Then, the sample contained in the tubes generate fluorescence by irradiation of light thus selected. The fluorescence thus generated, is reflected by reflection mirror(18) and focused through condensing lens(17). The fluorescence thus focused from each tubes is imaged on a light receiving element of the light receiving part to indicate and record the progress of reaction, continuously.
By the way, the light intensity generated from the light radiation lamp of the conventional apparatus, is differentiated depends on the positions, i.e., the center or the edges of the facet of light beam. Consequently, the light intensity generally varied with gaussian curve along with central axis of the facet of light beam and maintain this distribution even through the condensing lens.
This difference of brightness of light between the center and the edges of the facet of light beam, may cause a problem that the accurate data cannot be obtained since the fluorescence response from the sample positioned at the edges is weak. Various study and development have been tried to obviate this problem of the prior art. However, the technology which can overcome the above mentioned problem has not yet been suggested up to now. That is, a technology which can irradiate light with uniform intensity on the whole area of tube plate enlarged day by day, has not known in this field.
For the above reason, there is a critical limitation for enlarging the number of the tubes which can be monitored simultaneously. Therefore, the primary object of the present invention is to provide an apparatus for real-time monitoring the progress of biochemical reaction, which have technical part to irradiate light with uniform intensity over the whole area of the tube plate which is enlarged than that of prior art.
In addition, as depicted in FIG. 5, the reaction tube plate of prior art is in general rectangular shape. However the conventional light radiation system composed of a lamp and a lens, emits plane wave beam of which facet has round shape. Thus, some edge part of light beam should be eliminated to be fitted with the rectangular shaped tube plate. This problem lowers the efficiency of the source due to eliminated pat of light beam.
Various studies to obviate this problem of the low efficiency of light source have been suggested. However, any appropriate tools which can be adapted to a laboratory device integrated compactly, have not yet been developed.
Therefore, the present inventors had conceived that the sensitivity of the monitoring apparatus can be improved by strengthening the intensity of light by providing the light beam through waveguide which has a similar facet shape as that of the tube plate. Thus, another object of the claimed subject matter is to provide a real time monitoring apparatus for biochemical reaction which can irradiate light with uniform intensity over the whole area of the reaction tube plate, even over the whole area of the reaction tube plate enlarged than that of prior art, by irradiating light beam through a light irradiation part via a mirror.